Production of cephalexin via methoxymethyl ester

ABSTRACT

Cephalexin was produced by hydrolysis of its methoxymethyl ester which in turn was produced by reaction of 2-phenylglycine chloride hydrochloride with an imino ether of methoxymethyl 7phenoxyacetamido-3-methyl-ceph-3-em-4-carboxylate which had been prepared by reaction of an alcohol with the corresponding iminochloride made by treatment with PCl5 of methoxy-methyl 7phenoxyacetamido-3-methylceph-3-em-4-carboxylate. The last was made by thermal rearrangement of the methoxymethyl ester of phenoxymethylpenicillin sulfoxide.

United States Patent I19] Sapin0, Jr. et al.

[4 1 Oct. 22, 1974 [541 PRODUCTION OF CEPHALEXIN VIA METHOXYMETHYL ESTER both of NY.

[73] v Assignee: Bristol-Myers Company, New York,

[22] Filed: Feb. 8, 1973 21 Appl. No.: 330,770

[56] References Cited UNITED STATES PATENTS 3,632,578 l/l972 Chauvette 260/243 C 3,743,644 7/l973 Essery et al 260/243 C Primary Examiner-Nicholas S. Rizz'o Attorney, Agent, or Firm-Herbert W. Taylor, Jr.

[ 5 7] ABSTRACT Cephalexin was produced by hydrolysis of its methoxymethyl ester which in turn was produced by reaction of Z-phenylglycine chloride hydrochloride with an imino ether of methoxymethyl 7-phenoxyacetamido-3- methyl-ceph-3-em-4-carboxylate which had been prepared by reaction of an alcohol with the corresponding imino-chloride made by treatment with PCI of methoxy-methyl 7-phenoxyacetamido-3-methylceph- 3-em-4-carboxylate. The last was made by thermal rearrangement of the methoxymethyl ester of phenoxymethylpenicillin sulfoxide.

6 Claims, No Drawings PRODUCTION OF CEPHALEXIN VIA METHOXYMETHYL ESTER BACKGROUND OF THE INVENTION 7 and 3,676,429) which is then acylated to produce the N,N-diacyl-6-aminopenicillanic acid (as an easily hydrolyzed ester to protect the carboxyl group) from which the original acyl group of the starting penicillin is then removed along with the esterifying group, as by reaction with a nucleophile such as sodium thiophenolate, preferably in dimethylformamide. Such reactions were described in U.S. Pat. Nos. 3,594,367; 3,658,792 and 3,668,200 and in Farmdoc 14,183R, 48,2295 and 50,298S. Scientists of Yamanouchi Pharmaceutical Company, Ltd. have made related disclosures in patents abstracted as Farmdoc 08,653S, 40,656T, 47,171T and 47,172T and in Yakugaku Zasshi 92(4), 454-458 and 459464 (1972) which discloses the use of two particular esters of benzylpenicillin to make ampicillin.

Cephalexin is an antibacterial agent used in human therapy and marketed as the monohydrate of the free acid (he, the zwitterion). It is described, for example,

in J. Med. Chem. 12, 3103l3 (1969), J. Org. Chem. 36(9), 1259-1267 (1972) [See Belgium 765,596; Farmdoc 67,51lS] and in US. Pat. No. 3,507,861,

- U.K. 1,174,335 andCanada 856,786.

There are numerous disclosures of alternative methods for the production and purification of cephalexin (as the free acid). Examples include U.S. Pat. No. 3,634,416; 3,668,201; 3,668,202; 3,671,449 (Example 3,676,434; 3,676,437; 3,689,483. 3,694,437; 3,714,146; U.K. 1,227,014 and Belgium 768,306 (Farmdoc 80,0935) and by my colleagues J. Rubinfeld, R. U. Lemieux and R. Raap and U.S. application Ser. No. 143,683 filed May 11, 1971 (corresponding to Germany Off. 2,222,953 and Japan Publ. No. 42795/1972) and UK. 1,270,633.

Various disclosures with regard to crystalline forms and hydrates of cephalexin are given, for example, by

Pfeiffer et 211., in Journal of Pharmaceutical Sciences,

59(12), 1809-1814 (1970 by Bond et al. in Pharmaceutical Journal, 210-214 (August 22, 1970), in U.S. Pat. Nos. 3,502,663, 3,531,481; 3,665,656 and 3,692,781, in Belgium 753,910 (Farmdoc 8,2145) and Belgium 777,789 (Farmdoc 48,556T) and in France 2,096,l l7.

' The application of this transacylation processto cephalosporins developed more slowly and was complicated by the additional functionalgroups found in the only such compound produced (until recently) by fer- 11 g O c a ocn c-nii1T Iwsn; 2

. I coon I particular SUMMARY OF THE INVENTION 5 This invention comprises the useful intermediate methoxymethyl 3methyl-7-B-phenoxyacetamido-ceph-3- em-4-carboxylate and the process for the preparation of cephalexin which comprises the consecutive steps (which can be advantageously conducted in a single vessel) of a. reacting methoxymethyl 3-methyl-7-B- phenoxyacetamido-ceph-3-em-4-carboxylate in an an= hydrous, inert water-immiscible solvent preferably selected from the group consisting of methylene chloride, 1

dichloroethane, chloroform, tetrachloroethane, nitromethane and diethyl ether in the presence of an acid deactivating tertiary amine preferably selected from the group consisting of triethylamine, trimethylamine, dimethylaniline, quinoline, lutidine and pyridine with at least an equimolar amount of a halogenating agent preferably selected from the group consisting of phosphorus pentachloride, phosphorus pentabromide,

phosphorus trichloride, phosphorus tribromide, oxalyl chloride, p-toluenesulfonyl halide, phosphorus oxychloride and phosgene at a temperature in the range of 10 C. to -60 C. and preferably below 15" C.,

b. mixing therewith an aliphatic alcohol of less than seven carbons, preferably methanol, while maintaining the temperature in the range of 20 C. to -70 C. and preferably below 30 C., I

c. mixing therewith in portions at least an equimolar amount of D-(-)-2-phenylglycyl chloride hydrochloride below 30 C. and

d. mixing therewith water and a mineral acid, preferably sulfuric acid or hydrochloric acid, preferably below 10 C. to produce said cephalexin in the acidic, aqueous phase.

5 DETAILED DESCRIPTION OF THE INVENTION This invention provides a convenient synthetic route for the preparation of cephalexin from the methoxymethyl ester of 3desacetoxycephalosporin V which in turn is prepared byring expansion of penicillin V methoxymethyl ester sulfoxide (as described below or directly from 3desacetoxycephalosporin V acid and I chloromethyl methyl ether (as described below). This ester is also a useful intermediate for subsequent conversion by known side chain cleavage routes into 7- amlno-3desacetoxycephalosporanic acid (7-ADCA). Our new process renders unnecessary thepreparation of 7-ADCA'and enables one to simply exchange acyl group sidechains under essentially anhydrous conditions in good yield utilizing only one-reaction vessel.

o g 1 S 2H 6 5 CH +c H ocH :o ca

Q NaOH 0 H 4.5 DACVMI lE f /S C H OCH Ct IH-U +Pc1 +c H N(cH 0 Y CH3 RCH- (mew 1m) 2 2 h l 0 NH 0" 2 C1 +NaC1 I 5 C H OCH C=NU 3 2 O CH OCH 7 w 2 t 2 3 R represents phenyl.

0 (5on S 3 01 4 0 c n oca c=ug CO CH OCH a: The treatment of the methoxymethyl ester 1 with phosphorous pentachloride in methylene chloride solu- H N tion at 20 C., in the presence of N,N-dimethylaniline \Z 6 5 (CH3)2+RcHCOCl gave rise to the imidoyl chloride This intermediate was sufficiently stable that the reaction could easily be g followed to completion by thin layer chromatography.

Furthermore, in an analogous reaction sequence, using v phosphorous pentachloride in the presence of d pyridine in deuterochloroform solution at 25, it was a possible to characterize this intermediate fully by the use of NMR techniques. An NMR spectrum of the chlorination mixture in deuterochloroform, after '1 9 7 hour, revealed that the six line pattern between 5 and C H 0CH C 4 S g 6.3 ppm. (B-lactam; C C hydrogen atoms, coupled to N amide NH), initially present in the ester, was trans- CH formed into a well resolved pair of AB doublets (.l=4.5 f 3 Hz), characteristic of the chloroimide. Furthermore, NH Cl' CO CH OCH there was a downfield shift of the sidechain PhOCll 3 E 3 signal from 84.70 ppm. to 84.80 ppm., indicative of +C H NH CH (11' substitution of chlorine for oxygen at the amide car- 1; 6 5 3 2 bonyl group. These data clearly indicate the absence of any amide NH signal, which is generally submerged under the aromatic region center at 57.0 ppm. Finally, integration of the aromatic region revealed detection of only five protons due to 0,5,0, clearly showing the absence of amide NH signal. Peaks at 83.45 ppm. (C5 0) and 85.70 ppm. (CH OCH showed that the ester linkage was intact, and peaks at 52.10 ppm.

and 3.25 ppm (S-CHz) Were indicative of the dihydrothiazine moiety.

The chlorination mixture in methylene chloride solution was treated with excess methanol at to -30. After two hours, the complete disappearance of the chloroimide was observed by thin layer chromatography. The resulting imino-ether (3) was acylated with D-(-)-a-phenylglycyl chloride hydrochloride at '-20 to 30, giving rise to presumed intermediate,4. The acylation mixture was subsequently hydrolyzed in acidic water after ca. two hours, giving high quality cephalexin as a solid after pH adjustment in l-butanol/water to pH 4.5.

The structure of the cephalexin so produced was confirmed by elemental analysis, infrared, nuclear magnetic resonance and ultraviolet spectral techniques, as

well as TLC comparison with an authentic sample. Biopotency data indicated that the product was greater than 90 percent pure. Slightly lower quality cephalexin obtained during our preliminary investigations wasreadily recrystallized to analytically pure relatively water-insoluble monohydrate by pH adjustment in lbutanol/water at 25 in 70 percent recovery.

This invention is illustrated by the following examples, but it is to be understood that these examples are given by way of illustration and not of limitation. All temperatures are given in degrees Centigrade.

' DESCRIPTION OF THE PREFERRED EMBODIMENTS EXAMPLE I Conversion of Penicillin V to Desacetoxy cephalosporin V via ring expansion of Pen V sulfoxide methoxymethyl ester Preparation of Pen V sulfoxide methoxymethyl ester (PVSOMME: which is also named as methoxymethyl 6-phenoxyacetamido-penicillinate sulfoxide) After stirring 20 min. 30 ml. of chloromethyl methyl ether was added and the mixture stirred at 05 for 2 hr. The molecular sieves were removed by filtration and the solution was washed with two ml. portions of cold water. The organic layer was concentrated under reduced pressure to about 300 ml. and diluted to 1000 ml. with n-heptane. This gave a precipitate. The

slurry was stirred 2 hr. at 05, filtered and the result ing yellow solid redissolved in 500 ml. of 2-propanol at 40-50, diluted to 2000 ml. with heptane and cooled to 0-5 for 3 hr. The resulting solid was filtered, washed with 600 ml. of 50/50 2-propanol/heptane and dried at 35 for 18 hrs. There was obtained 107 g. (93

percent yield) of white, crystalline PVSOMME show- PVSOMME C-OCH OCH DA CV'MME RUN NO. 1

To 200 ml. of dioxane was added 20 gm. (0.050 mole) of PVSOMME, 10 g. of crushed (Linde type 4A). molecular sieves and 2.5 g. of the solid pyridine salt of p-toluenesulfonic acid. With stirring the mixture was heated to the reflux temperature for hours. After cooling and filtering, the mixture was concentrated under reduced pressure to an oil and taken up in 400 ml. dichloromethane. This solution was washed with three 200 ml. portions of water, carbon treated filtered and the filtrate dried and then concentrated under reduced pressure to give g. of a foamy solid. Thin layer chromatography showed the methoxymethyl ester of desacetoxy-ceph V (DACVMME) as the predominant product with a substantial amount of decarboxylated material and a trace of unchanged PVSOMME. This solid when subjected to cleavage conditions gave 7- ADCA assaying 72 percent pure by chemical assay in 32 percent yield.

RUN NO. 2.

The conditions of example l were repeated using 2 g. of pyridine tosylate salt and 4 hr. reaction time. After filtering and stripping, the methylene chloride solution was washed with 100 ml. of water, 100 ml. of pH 7 buffer solution, and 100 ml. of water. The dried organic solution on stripping gave 18 g. of yellow foamy solid. TLC showed about 65 percent DACVMME. A 100 mg.

sample was dissolved in 10 ml. of 50/50 acetone and pH 7 buffer solution and showed a potency of 6100 mcg./ml. in a DACV biological assay.

A 2 g. sample of this foamy solid was dissolved in 60 ml. of dichloromethane at 25 and treated 30 min. with l g. of 'meta-chloroperoxy-benzoic acid. The solution was washed with 30 ml. of water and 30 ml. of pH 7 buffer solution. The organic layer was dried, concentrated to an oil and the oil triturated with ether containing acetone. The resulting crystalline solid was filtered and dried to give 1.1 g. (52 percent yield) of desacetoxycephalosporin V sulfoxide methoxymethyl ester identical in all respects with authentic material.

RUN NO. 3

In a manner similar to the previous examples, 5 g. of PVSOMME, 50 ml. of dioxane, 7.5 g. of molecular sieves and 0.5 g. of pyridine salt of methanesulfonic acid was heated to reflux for 2 hrs. After filtering and stripping, the resulting oil was taken up in 50 ml. dichloromethane. This solution was washed twice with 30 ml. pH 7 buffer solution followed by 30 ml. cold water, carbon treated, dried and stripped to give 3.5 g. of light yellow foamy solid. On cleaving there was obtained 0.75 g. of 7-ADCA (27 percent yield) assaying 870 meg/mg. by chemical assay.

RUN NO. 4

To 10 g. of PYSOMME in 100 ml. of 1,2- dichloroethane was added 0.25 g. of pyridine tosylate and the mixture held at reflux temperature for 12 hours. TLC showed incomplete reaction. An additional 0.5 g. of catalyst was added and reflux continued for 4 hours. After cooling, the mixture was washed twice with 50 ml. of cold, acid (pH 1.5) water. The organic layer was dried and passed over a l X 60 cm. column of silica gel and the column eluted with 1250 ml. of dichloromethane. Appropriate fractions of the eluate were combined and stripped to give 2.0 g. (21 percent) of foamy solid identical with DACVMME.

RUNv NO. 5

The ring expansion was carried out on 5 g. PVSOMME in 50 ml. dioxane with 10 g. of crushed molecular sieves and 0.5 g. of pyridine tosylate at the reflux temp. for 18 hrs. After cooling and filtering 20 ml. of 6N HCl solution was added and the mixute held at about 25 approximately l-2 hours to hydrolyze the methoxy-methyl ester group. The mixture was then diluted with 50 ml. of water, extracted three times with 50 ml. of dichloromethane and the combined dichloromethane solution extracted three times with 50 ml. of pH 7 buffer solution. The combined buffer extracts were adjusted to pH 1.5 with HCl and extracted three times with 30 ml. of ethyl acetate. The combined ethylacetate layers were stripped to give a foamy solid which was taken up in 30 ml. of methanol and treated with dibenzylamine. After crystallizing 15 hrs. at 0, there was obtained 2.6 g. (48 percent yield) of the dibenzylamine salt of DACV identical with an authentic sample.

Methoxymethyl 3-methyl-7-B-phenoxyacetamidoceph- 3-em-4-carboxylate A suspension of 3-methyl-7-B- phenoxyacetamidoceph-3-em-4-carboxy1ic acid (20.0 g., 57.4 mmoles) (Netherlands 72/06193; Farmdoc 76,350T) in dry methylene chloride (300 ml.) was cooled to 05 and treated with triethylamine (7.96 ml., 5.80 g., 57.5 mmoles) in one portions under nitrogen. The resulting solution was then treated with chloromethyl methyl ether (5.0 ml., 5.03 g., 62.5 mmoles) over a 15 minute period, with high speed stirring at 05. The mixture was stirred for an additional 2.0 hours at 05. At this point, TLC on silica gel F 254, 250 p; system 4 acetone/ 12 benzene/ 1 HOAc revealed the absence of any starting acid at R 05, and the presence of the ester at R 075, when visualized with l percent potassium permanganate. The mixture was washed with 05 water (100 ml.), followed by 05 pH 7.00 phosphate buffer (100 ml.), dried over anhy-' drous sodium sulfate, and concentrated to an oil in vacuo at 20-25. The oil was dissolved in isopropanol (300 ml.) at 25, and seed crystals were added. The mixture was cooled to 05 and during the cooling period, the ester crystallized. The slurry was held at 05 for 4 hours, filtered, and the colorless cake displacement washed with 1:1 isopropanol/heptane (100 ml.). The colorless product was oven dried to constant weight at 40 giving 21.4 g. percent) of colorless rods of the title compound: m.p. 106l07; [a],,

+ 107 (g l, in methanol); A f" nm(loge) 215"(3.41), 255(3.l2), 260"'(3.15), 267(3.l6), 274(3.l2); TLC, silica gel F 254, 250 u, title compound R;0.75 (1 percent KMnO IR 1,800 (B-1actam C=O), 1718 (amide C=O), 3315 (NH) cm" in KBr; NMR (CDCl;,) 62.20(s,3,

Methoxymethyl 3-methyl-7-B- phenoxyacetamidoceph-3em-4-carboxylate (10.0 g., 25.4 mmoles) dissolved in dry methylene chloride (100 ml.) was cooled to 50. N,N-Dimethylaniline (7.45 ml., 7.15 g., 59.0 mmoles) was added, followed by the addition of phosphorous pentachloride (6.20 g., 29.77 mmoles) with high speed stirring. The mixture was brought to -30" to 35 and held at this temperature for 2 hours. At this point, thin layer chromatography on silica gel F 254, 250 1.; system 75 benzene/25 ethyl acetate showed the presence of the chloroimide at R, 0.8, and the absence of the starting material (R 0.5), when visualized with 1 percent potassium permanganate. The chlorination mixture was cooled to 50 and treated in one portion with 30 precooled dry methanol (30 ml., 23.7 g., 0.74 mole). The clear, light yellow solution was allowed to come to 35 and held at this temperature for 2 hours. At this point, thin layer chromatography in the above system revealed the complete disappearance of the chloroimide at R; 0.8. At -35, N,N-dimethylaniline (12.0 ml., 11.52 g., 95.07 mmole) was added and the mixture brought to -30" D-(-)-aphenylglycylchloride hydrochloride (5.8 g., 28.15 mmoles) was added'portion-wise over a 0.5 hour period at 30 with high speed stirring. Complete solution was evident within 5 minutes after the addition of acid chloride. The acylation mixture was held for 1 hour at 30 and gradually warmed to over a 0.5 hour period. Water (20 ml.) at 0-5was added with high speed stirring. The pH initially at 2.5 gradually dropped to 1.3 1.5 after 10 min. hydrolysis time. The pH was then adjusted to 0.3 0.5 with 6 N hydrochloric acid and allowed to proceed at 05 for 3 hours. At this point, thin layer chromatography on silica gel CF 254, 250 1.; system 50 acetone/l 5 benzene/1O HOAC/lS H O, visualized with 1 percent potassium permanganate revealed the presence of cephalexin at R, 0.8. The layers were separated and the aqueous layered with an equal volume of l-butanol. The mixture was adjusted to pH 4.5 with 30 percent aqueous sodium hydroxide at 05, and the mixture seeded immemdiately with cephalexin monohydrate. The mixture crystallized within 1 min. The slurry was held at 25 for 2 hours whereby the crystal structure changed from fine needles to large parallelograms. The mixture was cooled to 0-5 and held for 16 hours at pH 4.5. The product was collected, washed with cold water/ l-butanol, followed by l-butanol. The colorless product was oven dried at 40 to constant weight giving 3.5 g. (38 percent) of cephalexin monohydrate: TLC, silica gel F 254, 250 ;J., title compound at R, 0.8, system-50 acetone/1- 0HOAc/15H O (1% KMnO no other zones visible; [01],, 125.6 (Q 1, water); h nm(log e) 260(3.83), 2 l0(3.97); biopotency 888 ag/mg; IR 1770 (Blactam C=O), 1690 amide C=O), 1580 (CO{) cm; NMR (D O/d -DMSO/DC1) 62.09 (S, 3,

83.40(2d, 2,1 l 8 Hz, S-C1; l 85.05 (d, 1,.I-5Hz, C 5- lactam), 85.25(d, 2, J-5Hz, C B-lactam), 57.55 (S, 5, C655)- Anal. Calcd for C, H N 0,S-H O: C, 52.59; H, 5.24; N, 11.50; S. 8.73; KF, 4.93.

Found: C, 52.49; H, 5.17; N, 10.96; S, 8.73; KF, 5.62.

Recrystallization of Cephalexin Monohydrate Cephalexin monohydrate (1.0 g.) was dissolved in 1:1 water/l-butanol by adjusting the pH to 1.0 with 6 Nhydrochloric acid at 25. The mixture was warmed to 35-40 and slowly adjusted to pH 4.5 with 10 percent aqueous sodium hydroxide wi'th seeding at pH 3.0. The mixture was crystallized for 30 min. at 35 and cooled to 25 over a 45 min. period and held at this temperature for 1 hr. The slurry was cooled to 05 and held at this temperature for 16 hours. The product was collected, washed with 05 1:1 water/l-butanol, l-butanol and oven dried to constant weight at 40 giving 700 mg. (70 percent recovery), identical in all respects with that obtained in the acylation sequence above. The biopotency ranged between 915 ug/mg and 1065 ug/mg.

EXAMPLE 2 3-Methyl-7-B-[D-2amino-2-phenylacetamido]ceph-3- em-4-carboxylate (cephalexin). Methoxymethyl-3methyl-7-B- phenoxyacetamidoceph-3-em-4-ca'rboxylate (5.0 g., 12.7 mmoles) was dissolved in dry methylene chloride (50 ml.) and cooled to 50. N,N-Dimethylaniline (3.72 ml., 3.58 g., 29.5 mmoles) was added and the mixture treated with phosphorous pentachloride (3.1 g., 14.9 mmoles). The mixture was held at 30 to -35 for 2 hours and cooled to 60. The mixture was treated in one portion with dry, precooled (-60) methanol (20 ml., 15.8 g., 0.49 mole) and the mixture brought to 35 to 40. After 2 hours at this temperature, N,N-dimethylaniline (6.0 ml., 5.76 g., 47.53 mmoles) was added, followed by the portionwise addition of D-(-)-a-phenylglycyl chloride hydrochloride (2.9 g., 14.08 mmoles) over a 0.5 hour period. The.

mixture was held at 35 for 1 hour, water (30 ml.) was added, and the mixture stirred at 05 for 10 min., and 1 hour at 25. The layers were separated and the aqueous concentrated to ca. 15-18 ml. Acetonitrile (20 ml.) was added, and the pH adjusted to 4.5 5.0 with 20 percent aqueous sodium hydroxide at 05. Cephalexin began to crystallize within l minute. The slurry was held at 05 for 6 hours, the product collected, washed with water, and acetonitrile and oven dried at 40 giving 1.75 g. (38 percent) of pale yellow cephalexin: TLC, silica gel F 254, 250 a, title compound R 0.80, system50 acetonelHOAc/benzene/lO HoAc/ 15 H 0 (1 percent KMnOi), no other Zones were visible; NMR (60 MHz), showed traces of acetonitrile. The above product was recrystallized by pH adjustment in l-butanol/water as described previously in 69 percent recovery which was identical in all respects with that obtained previously.

EXAMPLE 3 3-Methyl-7-fi-[ D'2-amino-2phenylacetamido ]-ceph- 3-em4-carboxylate (cephalexin).

Methoxymethyl 3-methyl-7-B-phenoxyacetamidoceph-3-em-4-carboxylate (10.0 g., 25.4 mmoles) dissolved in dry methylene chloride ml.) was cooled to -20. N,N-dimethylaniline (7.45 ml., 7.15 g., 59.0 mmoles) was added followed by the addition of phosphorouspentachloride (6.20 g., 29.77 mmoles) with high speed stirring. The mixture was held at 20 for 2 hours, and treated with precooled (-30) methanol (30 ml., 23.7 g., 0.74 mole). The mixture was brought to 20 and held for 2 hours at this temperature. D-(-)-or-Phenyl-glycylchloride hydrochloride (5.8 g.,

28.15 mmoles) was added portionwise over a 0.5 hour period at 20 with high speed stirring. The acylation was allowed to proceed for 1 hour at -20 and gradually brought to over a 1 hour period. Water ml.) was added and the mixture brought to The pH initially at 2.5 dropped to 1.8 after 10-15 min. hydrolysis time. The pH was then adjusted to 0.2 with 6 111 HCl and the mixture stirred at 25 for 3 hours. The layers were separated and the aqueous layered with an equal volume of l-butanol. The mixture was adjusted to pH 4.5 over a min. period with seeding at pH 3.0. The mixture crystallized within 2 minutes at pH 4. The slurry was allowed to crystallize at 25 for 2 hours and then cooled to 0-5 and held for 16 hours. The product was collected and washed with 1:1 water/ 1 butanol, butanol, isopropanol, and oven dried at 45 giving 3.70 g. (40.0 percent) of the title compound which was identical in all respects with that obtained earlier. A second crop of slightly less pure cephalexin was obtained by evaporation of the mother liquor above and cooling for 3 hours at 0-5 giving 1.20 g. (13 percent) for a total yield of 53 percent.

1R data were obtained on a Beckmann 1R9. NMR spectra were recorded on a Perkin Elmer Rl2-B operating at 60MHz with probe temperature at ca. 40. NMR data are given in units relative to tetramethylsilane. U.V. data were obtained on a Beckmann Acta 3. Rotation data given were obtained at 25 on a Perkin Elmer PEl4l recording polarimeter. The superscript refers to a shoulder on the UV curve. Bioassay data were obtained by turbidimetric methods using S. Au-

reus 209P.

We claim:

1. The process for the preparation of cephalexin which comprises the consecutive steps of a. reacting methoxymethyl 3-methyl-7-B- phenoxyacetamido-ceph-3-em-4-carboxylate in an anhydrous, inert water-immiscible solvent in the presence of an acid deactivating tertiary amine with at least an equimolar amount of a halogenating agent at a temperature in the range of 10 C.

to 60 C.

b. mixing therewith an aliphatic alcohol of less than seven carbons while maintaining the temperature in the range of 20 C. to 70 C.

c. mixing therewith in portions at least an equimolar amount of D-(-)-2-phenylglycyl chloride hydrochloride below 30 C. and

d. mixing therewith water and a mineral acid to produce said cephalexin in the acidic, aqueous phase.

2. The process of claim 1 for the preparation of cephalexin which comprises the consecutive steps of a. reacting methoxymethyl 3-methyl-7-B- phenoxyacetamido-ceph-3-em-4-carboxylate in an anhydrous, inert water-immiscible solvent in the presence of an acid deactivating tertiary amine V with at least an equimolar amount of a halogenating agent selected from the group consisting of phosphorus pentachloride, phosphorus pentabromide, phosphorus trichloride, phosphorus tribromide, oxalyl chloride, p-toluenesulfonyl halide, phosphorus oxychloride and phosgene at a temperature in the range of 10 C. to 60 C.

b. mixing therewith an aliphatic alcohol of less than seven carbons while maintaining the temperature in the range of 20 C. to 70 C.

LII

c. mixing therewith in portions at least an equimolar amount of D-(-)-2-phenylglycyl chloride hydrochloride below 30 C. and

d. mixing therewith water and a mineral acid to produce said cephalexin in the acidic, aqueous phase.

3. The process of claim 1 for the preparation of cephalexin which comprises the consecutive steps of a. reacting methoxymethyl 3-methyl-7-B- phenoxyacetamido-ceph-3-em-4-carboxy1ate in an anhydrous, inert water-immiscible solvent selected from the group consisting of methylene chloride, dichloroethane, chloroform, tetrachloroethane, nitromethane and diethyl ether in the presence of an acid deactivating tertiary amine selected from the group consisting of triethylamine, trimethylamine, dimethylaniline, quinoline, lutidine and pyridine with at least an equimolar amount of a halogenating agent selected from the group consisting of phosphorus pentachloride, phosphorus pentabromide, phosphorus trichloride, phosphorus tribromide, oxalyl chloride, p-toluene-sulfonyl halide, phosphorus oxychloride and phosgene at a temperature in the range of -l0 C. to 60 C.

b. mixing therewith an aliphatic alcohol of less than seven carbons while maintaining the temperature in the range of 20 C. to C.

c. mixing therewith in portions at least an equimolar amount of D-(-)-2-phenylglycyl chloride hydrochloride below 30 C. and

d. mixing therewith water and a mineral acid to produce said cephalexin in the acidic, aqueous phase.

4. The process of claim 1 for the preparation of cephalexin which comprises the consecutive steps of a. reacting methoxymethyl 3-methyl-7-B- phenoxyacetamido-ceph-3-em-4-carboxylate in an anhydrous, inert water-immiscible solvent selected from the group consisting of methylene chloride, dichloroethane, chloroform, tetrachloroethane, nitromethane and diethyl ether in the presence of an acid deactivating tertiary amine selected from the group consisting of triethylamine, trimethylamine, dimethylaniline, quinoline, lutidine and pyridine with at least an equimolar amount of phosphorus pentachloride at a temperature in the range of 10 C. to 60 C.

b. mixing therewith an aliphatic alcohol of less than seven carbons while maintaining the temperature in the range of 20 C. to 70 C.

c. mixing therewith in portions at least an equimolar amount of D-(-)-2-phenylglycyl chloride hydrochloride below -30 C. and

d. mixing therewith water and a mineral acid to produce said cephalexin in the acidic, aqueous phase.

5. The process of claim 1 for the preparation of cephalexin which comprises the consecutive steps of a. reacting methoxymethyl 3-methyl-7-B- phenoxyacetamido-ceph-3em-4-carboxylate in an anhydrous, inert water-immiscible solvent selected from the group consisting of methylene chloride, dichloroethane, chloroform, tetrachloroethane, nitromethane and diethyl ether in the presence of an acid deactivating tertiary amine selected from the group consisting of triethylamine, trimethylamine, dimethylaniline, quinoline, lutidine and pyridine 13 14 with at least an equimolar amount of phosphorus methylene chloride in the presence of dimethylanpentachloride at a temperature below 1 C., iline with at least an equimolar amount of phosphob. mixing therewith methanol while maintaining the [Us hl id at a temperature b l C temperature below 30 C.,

c. mixing therewith in portions at least an equimolar 5 amount of D-(J-Z-phenylglycyl chloride hydrochloride below 30 C. and

d. mixing therewith water and a mineral acid to produce said cephalexin in the acidic, aqueous phase.

b. mixing therewith methanol while maintaining the temperature below -30 C.,

c. mixing therewith in portions at least an equimolar amount of D-( -)-2-phenylglycyl chloride hydrol0 chloride below 30 C. and

6. The process of claim 1 for the preparation of cephmixing therewith Water and hlydl'ochlorlc aCld to alexin which comprises the consecutive steps of Produce Said cephalexirl in the acidic aqueous a. reacting methoxymethyl 3-methyl-7-B- phase.

phenoxyacetamido-ceph-3-em-4-carboxylate in 

1. THE PROCESS FOR THE PREPARATION OF CEPHALEXIN WHICH COMPRISES THE CONSECUTIVE STEPS OF A. REACTING METHOXYMETHYL 3-METHYL-7-BPHENOXYACETAMIDO-CEPH-3-EM-4-CARBOXYLATE IN AN ANHYDROUS, INERT WATER-IMMISCIBLE SOLVENT IN THE PRESENCE OF AN ACID DEACTIVATING TETIARY AMINE WITH AT LEAST AN EQUIMOLAR AMOUNT OF A HYDROGENATING AGENT AT A TEMPERATURE IN THE RANGE OF -10*C. TO -60*C. B. MIXING THEREWITH AN ALIPHATIC ALCOHOL OF LESS THAN SEVEN CARBONS WHILE MAINTAINING THE TEMPERATURE IN THE RANGE OF -20*C. TO -70*C. C. MIXING THEREWITH IN PORTIONS AT LEAST AN EQUIMOLAR AMOUNT OF D-(-)-2-PHENYLGLYCYL CHLORRIDE HYDROCHLORIDE BELOW -30:C. AND D. MIXING THEREWITH WATER AND A MINERAL ACID TO PRODUCE SAID CEPHALEXIN IN THE ACIDIC, AQUEOUS PHASE.
 2. The process of claim 1 for the preparation of cephalexin which comprises the consecutive steps of a. reacting methoxymethyl 3-methyl-7- Beta -phenoxyacetamido-ceph-3-em-4-carboxylate in an anhydrous, inert water-immiscible solvent in the presence of an acid deactivating tertiary amine with at least an equimolar amount of a halogenating agent selected from the group consisting of phosphorus pentachloride, phosphorus pentabromide, phosphorus trichloride, phosphorus tribromide, oxalyl chloride, p-toluenesulfonyl halide, phosphorus oxychloride and phosgene at a temperature in the range of -10* C. to -60* C. b. mixing therewith an aliphatic alcohol of less than seven carbons while maintaining the temperature in the range of -20* C. to -70* C. c. mixing therewith in portions at least an equimolar amount of D-(-)-2-phenylglycyl chloride hydrochloride below -30* C. and d. mixing therewith water and a mineral acid to produce said cephalexin in the acidic, aqueous phase.
 3. The process of claim 1 for the preparation of cephalexin which comprises the consecutive steps of a. reacting methoxymethyl 3-methyl-7- Beta -phenoxyacetamido-ceph-3-em-4-carboxylate in an anhydrous, inert water-immiscible solvent selected from the group consisting of methylene chloride, dichloroethane, chloroform, tetrachloroethane, nitromethane and diethyl ether in the presence of an acid deactivating tertiary amine selected from the group consisting of triethylamine, trimethylamine, dimethylaniline, quinoline, lutidine and pyridine with at least an equimolar amount of a halogenating agent selected from the group consisting of phosphorus pentachloride, phosphorus pentabromide, phosphorus trichloride, phosphorus tribromide, oxalyl chloride, p-toluene-sulfonyl halide, phosphorus oxychloride and phosgene at a temperature in the range of -10* C. to -60* C. b. mixing therewith an aliphatic alcohol of less than seven carbons while maintaining the temperature in the range of -20* C. to -70* C. c. mixing therewith in portions at least an equimolar amount of D-(-)-2-phenylglycyl chloride hydrochloride below -30* C. and d. mixing therewith water and a mineral acid to produce said cephalexin in the acidic, aqueous phase.
 4. The process of claim 1 for the preparation of cephalexin which comprises the consecutive steps of a. reacting methoxymethyl 3-methyl-7- Beta -phenoxyacetamido-ceph-3-em-4-carboxylate in an anhydrous, inert water-immiscible solvent selected from the group consisting of methylene chloride, dichloroethane, chloroform, tetrachloroethane, nitromethane and diethyl ether in the presence of an acid deactivating tertiary amine selected from the group consisting of triethylamine, trimethylamine, dimethylaniline, quinoline, lutidine and pyridine with at least an equimolar amount of phosphorus pentachloride at a temperature in the range of -10* C. to -60* C. b. mixing therewith an aliphatic alcohol of less than seven carbons while maintaining the temperature in the range of -20* C. to -70* C. c. mixing therewith in portions at least an equimolar amount of D-(-)-2-phenylglycyl chloride hydrochloride below -30* C. and d. mixing thereWith water and a mineral acid to produce said cephalexin in the acidic, aqueous phase.
 5. The process of claim 1 for the preparation of cephalexin which comprises the consecutive steps of a. reacting methoxymethyl 3-methyl-7- Beta -phenoxyacetamido-ceph-3-em-4-carboxylate in an anhydrous, inert water-immiscible solvent selected from the group consisting of methylene chloride, dichloroethane, chloroform, tetrachloroethane, nitromethane and diethyl ether in the presence of an acid deactivating tertiary amine selected from the group consisting of triethylamine, trimethylamine, dimethylaniline, quinoline, lutidine and pyridine with at least an equimolar amount of phosphorus pentachloride at a temperature below -15* C., b. mixing therewith methanol while maintaining the temperature below -30* C., c. mixing therewith in portions at least an equimolar amount of D-(-)-2-phenylglycyl chloride hydrochloride below -30* C. and d. mixing therewith water and a mineral acid to produce said cephalexin in the acidic, aqueous phase.
 6. The process of claim 1 for the preparation of cephalexin which comprises the consecutive steps of a. reacting methoxymethyl 3-methyl-7- Beta -phenoxyacetamido-ceph-3-em-4-carboxylate in methylene chloride in the presence of dimethylaniline with at least an equimolar amount of phosphorus pentachloride at a temperature below -15* C., b. mixing therewith methanol while maintaining the temperature below -30* C., c. mixing therewith in portions at least an equimolar amount of D-(-)-2-phenylglycyl chloride hydrochloride below -30* C. and d. mixing therewith water and hydrochloric acid to produce said cephalexin in the acidic aqueous phase. 